• Title/Summary/Keyword: p40 subunit

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Characterization of Polyphenoloxidase extracted from Solanum tuberosum Jasim

  • Jang, Jae-Won;Ma, Yu-Hyun;Shin, Ju-Mi;Song, Kyung-Bin
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.117-122
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    • 2005
  • Polyphenoloxidase (PPO) was extracted from Solanum tuberosum Jasim by various chromatographic methods and was subsequently purified and characterized. PPO was purified upto 78-fold from the crude extract. SDS-PAGE profile of the enzyme showed a major subunit of PPO with molecular weight of 40 kDa. The optimum pH and temperature for the maximum activity of PPO was 6.5 and $25^{\circ}C$, respectively. The enzyme was found to be quite stable between 10 and $40^{\circ}C$, whereas it was almost inactivated at $70^{\circ}C$ when incubated for 30 min. Substrate specificity study indicated that catechol was the most suitable substrate for PPO isolated from purple-fleshed potato with a $K_m$ value of 21.1 mM. The most effective inhibitor was ascorbic acid, followed by L-cysteine, citric acid, EDTA, and boric acid. Studies on the effect of metal ion on PPO activity showed that magnesium and copper were inhibitory, while iron and zinc ions increased the activity of PPO.

A Phi Class Glutathione S-transferase from Oryza sativa (OsGSTF5): Molecular Cloning, Expression and Biochemical Characteristics

  • Cho, Hyun-Young;Lee, Hae-Joo;Kong, Kwang-Hoon
    • BMB Reports
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    • v.40 no.4
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    • pp.511-516
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    • 2007
  • A glutathione S-transferase (GST) related to the phi (F) class of enzymes only found in plants has been cloned from the Oryza sativa. The GST cDNA was cloned by PCR using oligonucleotide primers based on the OsGSTF5 (GenBank Accession No. $\underline{AF309382}$) sequences. The cDNA was composed of a 669-bp open reading frame encoding for 223 amino acids. The deduced peptide of this gene shared on overall identity of 75% with other known phi class GST sequences. On the other hands, the OsGSTF5 sequence showed only 34% identity with the sequence of the OsGSTF3 cloned by our previous study (Cho et al., 2005). This gene was expressed in Escherichia coli with the pET vector system and the gene product was purified to homogeneity by GSH-Sepharose affinity column chromatography. The expressed OsGSTF5 formed a homo-dimer composed of 28 kDa subunit and its pI value was approximately 7.8. The expressed OsGSTF5 displayed glutathione conjugation activity toward 1-chloro-2,4-dinitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy)propane and glutathione peroxidase activity toward cumene hydroperoxide. The OsGSTF5 also had high activities towards the herbicides alachlor, atrazine and metolachlor. The OsGSTF5 was highly sensitive to inhibition by S-hexylGSH, benastatin A and hematin. We propose from these results that the expressed OsGSTF5 is a phi class GST and appears to play a role in the conjugation of herbicide and GPOX activity.

Characterization of Isocitrate Lyase from Micrococcus luteus (Micrococcus luteus에서 정제한 Isocitrate Lyase의 특성)

  • 정기택;서승교;우철주;박임동;정병태;박영호
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.230-236
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    • 1993
  • The isocitrate lyase extracted from Micrococcus luteus was purified 38.8 folds with the overall yield of 10.2%, by the ammonium sulfate fractionation, DEAE-cellulose, 1st Sephadex G-200 and 2nd Sephadex G-200 column chromatography. The purified enzyme showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified enzyme was estimated 60,000 by the SDS-polyacry]amide gel electrophoresis. The apparent Michaelis constant, Km value for isocitrate was 0.95 mM. The optimum pH and temperature of the purified enzyme were pH 7.5 and $40^{\circ}C$, respectively. The enzyme was activated by $Mg^{2+}$ and inhibited by $Mn^{2+}$, $Ca^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $CO^{2+}$. In addition, the activity of isocitrate lyase was increased by glutathione and 2-mercaptocthanol at 5 mM and cysteine at I mM.

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Purification and Characterization of the Anabolic Acetolactate Synthase III from Serratia marcescens ATCC 25419

  • Joo, Han-Seung;Kim, Soung-Soo
    • BMB Reports
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    • v.34 no.3
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    • pp.244-249
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    • 2001
  • The anabolic acetolactate synthase III was purified to homogeneity from Serratia marcescens using DEAE-Sepharose, Phenyl-Sepharose, and hydroxylapatite column chromatography The native molecular weight of the enzyme was approximately 165 kDa. The enzyme is composed of two large and two small subunits with molecular weights of 64 and 15 kDa, respectively. The N-terminal sequence of the large and small subunit of the enzyme was Ser-Ala-Thr-Pro-Gln-Pro-Ser-Thr-Arg-Phe-Thr-Cys-Ala-Gln-Leu-Ile-Ala-His-Leu and Met-Leu-Gln-Pro-Gln-Asp-Lys-Pro-Gln-Val-Ile-Leu-Glu-Leu-Ala-Val-Arg-Asn-His-Pro-Gly-Val-Met-Ser-His-Val, respectively. The optimum pH and pI value were 7.5 and 5.5, respectively The $IC_{50}$ values were $20\;{\mu}M$ and $14\;{\mu}M$ for valine and herbicide SU7, respectively. The substrate specificity ratio, R value, was determined to be approximately 40, which suggests that this enzyme prefers the formation of $\alpha$-aceto-$\alpha$-hydroxybutyrate leading to the synthesis of isoleucine.

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Three-dimensional analysis of the anterior loop of the inferior alveolar nerve in relation to the growth pattern of the mandibular functional subunit

  • Yoon, Seungkyu;Kim, Jae-Young;Jeong, Cheol-Hee;Park, Jengbin;Huh, Jong-Ki;Park, Kwang-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.40
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    • pp.30.1-30.6
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    • 2018
  • Background: The purpose of the present study was to investigate the differences in the position and shape of the anterior loop of the inferior alveolar nerve (ALIAN) in relation to the growth pattern of the mandibular functional subunit. Methods: The study was conducted on 56 patients among those who had undergone orthognathic surgery at the Gangnam Severance Hospital between January 2010 and December 2015. Preoperative computerized tomography (CT) images were analyzed using the Simplant OMS software (ver.14.0 Materialise Medical, Leuven, Belgium). The anterior and inferior lengths of ALIAN (dAnt and dInf) and each length of the mandibular functional subunits were measured. The relationship between dAnt, dInf, and the growth pattern of the mandibular subunits was analyzed. Results: The length of the anterior portion of ALIAN (dAnt) reached 3.34 ± 1.59 mm in prognathism and 1.00 ± 0.97 mm in retrognathism. The length of the inferior portion of ALIAN (dInf) reached 6.81 ± 1.33 mm in prognathism and 5.56 ± 1.34 mm in retrognathism. The analysis of Pearson's correlation coefficiency on all samples showed that the lengths of functional subunits were positively correlated with the loop depth. The length of the symphysis area in prognathic patients was positively correlated with the anterior loop depth (p = 0.005). Conclusions: Both the anterior and inferior length of ALIAN are longer in prognathic patients. Especially, it seems to be associated with the growth of the symphysis area.

Purification and Characterization of an Alkaline Protease Produced by Alkalophilic Bacillus sp. DK1122 (호알칼리성 Bacillus sp. DK1122 균주가 생산하는 알칼리성 단백질 분해효소의 정제 및 특성)

  • Lee, Hyungjae;Yoo, Ji-Seung;Bai, Dong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.333-340
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    • 2016
  • An alkaline protease was purified and characterized from an alkalophilic microorganism, Bacillus sp. DK1122, isolated from soil in central Korea. The optimum temperature and pH for the growth of the producer strain were 40℃ and pH 9.0, respectively. The protease was produced aerobically at 40℃ after 24 h incubation in modified Horikoshi I medium (pH 9.0) containing 0.5% (w/v) glucose, 0.8% (w/v) yeast extract, 0.5% (w/v) polypeptone, 0.1% (w/v) K2HPO4, 0.02% (w/v) MgSO4·7H2O, 1% (w/v) Na2CO3, and 3% (w/v) NaCl. The alkaline protease was purified by 70% ammonium sulfate precipitation of the culture supernatant of Bacillus sp. DK1122, followed by CM-Sepharose chromatography. The molecular weight of the enzyme was estimated to be 27 kDa on the basis of SDS-PAGE. The optimum temperature and pH for the protease activity were 60℃ and pH 9.0, respectively. Addition of CaCl2 increased the thermal stability of the purified protease, where 90% of protease activity was retained at 60℃ for up to 3 h. Consequently, it is expected that the alkaline protease from this study, exhibiting stability at pH 7–9 and 60℃, may be promising for application in the food and detergent industries.

Effects of SNP in TSH-β Gene of Chicken on Economic Traits (닭의 경제 형질에 미치는 TSH-β 유전자 변이 효과 분석)

  • Seo, Joohee;Oh, Jae Don;Choi, Eun Ji;Lim, Hee Kyong;Seong, Jiyeon;Song, Ki Duk;Lee, Jun Heon;Lee, Hak Kyo;Kong, Hong Sik;Jeon, Gwang Joo;Shon, Young Gon;Choi, Kang Duk
    • Korean Journal of Poultry Science
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    • v.40 no.2
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    • pp.115-120
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    • 2013
  • Thyroid hormone (TH) plays a role in growth of the poultry. Thyroid-stimulating hormone (TSH) stimulates production and distribution of TH, and is a heterodimer which is formed by ${\alpha}$- and ${\beta}$-subunits. Most of TSH activity is known to rely on ${\beta}$-subunit. TSH-${\beta}$ gene is located on chicken chromosome 26 and associated with growth performances. Therefore, this study aimed to investigate the association of TSH- ${\beta}$ SNP (G1031C) with economic traits (layday, layw, layno, bw150, bw270, layw270) in Korean Native Black chicken, Rhode Island Red and Cornish. Allele frequency of GG genotype in Rhode Island Red (RIR) was found to be 1.00 in this study. A significant effect was only observed on body weight at day 150 in Cornish. In Cornish, body weights of chicken with the CC genotype ($302.15{\pm}6.336$) were significantly higher than that of the GG genotype ($294.56{\pm}4.537$) (p<0.05). These findings suggest that the G1031C SNP of TSH-${\beta}$ gene can be used for improvement of growth-related traits in Cornish.

Characterization of Extracellular Peroxidase from Pleurotus ostreatus (Pleurotus ostreatus에서 분비되는 Peroxidase의 특성)

  • 배성호;신광수;강사욱;하영칠;최선진;김규중;최형태
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.348-356
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    • 1989
  • An extracellular peroxidase found in culture broth of Pleurotus ostreatus was induced by syringic acid. This enzyme was fractionated by DEAE Sephadex A-50 ion exchange chromatography and gel filtration chromatogrphy on Sephadex G-150. The enzyme is a glycoprotein containing 35.7% carbohydrate. The results of SDS-linear polyacrylamide gradient gel electrophoresis and gel filtration indicate that the enzyme is a dimer consisted of identical subunits (Mr=72,400). The absorption spectrum of the enzyme indicates the presence of one mole of iron protoporphyrin IX per one mole of subunit. Isoelectric point of the enzyme is 4.26 and $K_m$ values for $H_2O_2$ is $7.2{\mu}M$. The enzyme showed its optimal activity at pH 3.5-4.0 and at $40^{\circ}C$. The Km values of this enzyme for ferulic acid and sinapic acid are 2.4 and 12.3 times higher than those of horseradish peroxidase, respectively.

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Relationships between Single Nucleotide Polymorphism Markers and Meat Quality Traits of Duroc Breeding Stocks in Korea

  • Choi, J.S.;Jin, S.K.;Jeong, Y.H.;Jung, Y.C.;Jung, J.H.;Shim, K.S.;Choi, Y.I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.9
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    • pp.1229-1238
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    • 2016
  • This study was conducted to determine the relationships of five intragenic single nucleotide polymorphism (SNP) markers (protein kinase adenosine monophosphate-activated ${\gamma}3$ subunit [PRKAG3], fatty acid synthase [FASN], calpastatin [CAST], high mobility group AT-hook 1 [HMGA1], and melanocortin-4 receptor [MC4R]) and meat quality traits of Duroc breeding stocks in Korea. A total of 200 purebred Duroc gilts from 8 sires and 40 dams at 4 pig breeding farms from 2010 to 2011 reaching market weight (110 kg) were slaughtered and their carcasses were chilled overnight. Longissimus dorsi muscles were removed from the carcass after 24 h of slaughter and used to determine pork properties including carcass weight, backfat thickness, moisture, intramuscular fat, $pH_{24h}$, shear force, redness, texture, and fatty acid composition. The PRKAG3, FASN, CAST, and MC4R gene SNPs were significantly associated with the meat quality traits (p<0.003). The meats of PRKAG3 (A 0.024/G 0.976) AA genotype had higher pH, redness and texture than those from PRKAG3 GG genotype. Meats of FASN (C 0.301/A 0.699) AA genotype had higher backfat thickness, texture, stearic acid, oleic acid and polyunsaturated fatty acid than FASN CC genotype. While the carcasses of CAST (A 0.373/G 0.627) AA genotype had thicker backfat, and lower shear force, palmitoleic acid and oleic acid content, they had higher stearic acid content than those from the CAST GG genotype. The MC4R (G 0.208/A 0.792) AA genotype were involved in increasing backfat thickness, carcass weight, moisture and saturated fatty acid content, and decreasing unsaturated fatty acid content in Duroc meat. These results indicated that the five SNP markers tested can be a help to select Duroc breed to improve carcass and meat quality properties in crossbred pigs.

Salubrinal Alleviates Pressure Overload-Induced Cardiac Hypertrophy by Inhibiting Endoplasmic Reticulum Stress Pathway

  • Rani, Shilpa;Sreenivasaiah, Pradeep Kumar;Cho, Chunghee;Kim, Do Han
    • Molecules and Cells
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    • v.40 no.1
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    • pp.66-72
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    • 2017
  • Pathological hypertrophy of the heart is closely associated with endoplasmic reticulum stress (ERS), leading to maladaptations such as myocardial fibrosis, induction of apoptosis, and cardiac dysfunctions. Salubrinal is a known selective inhibitor of protein phosphatase 1 (PP1) complex involving dephosphorylation of phospho-eukaryotic translation initiation factor 2 subunit $(p-eIF2)-{\alpha}$, the key signaling process in the ERS pathway. In this study, the effects of salubrinal were examined on cardiac hypertrophy using the mouse model of transverse aortic constriction (TAC) and cell model of neonatal rat ventricular myocytes (NRVMs). Treatment of TAC-induced mice with salubrinal ($0.5mg{\cdot}kg^{-1}{\cdot}day^{-1}$) alleviated cardiac hypertrophy and tissue fibrosis. Salubrinal also alleviated hypertrophic growth in endothelin 1 (ET1)-treated NRVMs. Therefore, the present results suggest that salubrinal may be a potentially efficacious drug for treating pathological cardiac remodeling.